Difference between revisions of "Team:Aix-Marseille/Notebook"

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===== Tuesday, September 6th =====
 
===== Tuesday, September 6th =====
  
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* Transformation of cadA mutant competant strain by Bba_K1951004 in pSB1C3, spread on Kana/cm petri dishes
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* Transformation of cadA mutant competant strain by Bba_K1951004 in pSB1C3, spread on Kana/Cm petri dishes
 
* Colony PCR Bba_K1951004 in pSB1C3
 
* Colony PCR Bba_K1951004 in pSB1C3
 
* Starters from Bba_K1951004 in pSB1C3
 
* Starters from Bba_K1951004 in pSB1C3
 
* 1 step of the transduction using phage p1 from fliC mutant Keio bank. Conserved at 4°C with 20µL chlorophorme
 
* 1 step of the transduction using phage p1 from fliC mutant Keio bank. Conserved at 4°C with 20µL chlorophorme
 +
* PCR with Q5 master mix using oligo of diriged mutagenesis for the insertion of Bbs1 site in the 214-215 and 238-239 sites of BBa_K1951008. Digest 3h with addition of 1µL DpNI in the PCR tube.
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* Transformation of the previous PCR in competant Tg1
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* Starters of the PCR from 09.02.2016 
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* Remade PCR on Bba_K1951001(DesB), Bba_K1951002(DesC), Bba_K1951003(DesD) with Bba_B0034 (rbs) + BBa_I0500(Prom) but still no result
 
</div>
 
</div>
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<div id="2016-09-07" class="evday">
  
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===== Wednesday, September 7th =====
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 +
* Colony PCR on cadA mutant complemented with BBa_K1951004
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* HPLC test of cadA mutant complemented with BBa_K1951004
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* Colony PCR to verify the insertion of Bbs1 site in the 214-215 and 238-239 sites of BBa_K1951008 and BBa_K1951009
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* From a starter of intermediate biobrick BBa_K1951004+ RBS BBa_K1951001, made culture at DO=0.2 until DO=0.6 and took a sample of 1uDO. Added 50µL beta mercatoethanol to the sample and congeled at -20°C to test protein production on a SDS page/Comassie
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 +
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</div>
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<div id="2016-09-08" class="evday">
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===== Thursday, September 8th =====
 +
 +
* Starters of BBa_K1951008 + BbsI 215-216 position and starters of BBa_K1951008 + BbsI 238-239 position
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* Spreading of transduction in a <i> E. coli W3110</i> strain using the phage P1 from the 09.05.2016 (500£µL cells + 2µL P1 or 10µL P1 or 10µL P1 or 50µL P1 or 100µL P1)
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* Digestion of intermediate biobrick BBa_K1951004+ RBS BBa_K1951001 by EcoRI and SpeI and RBS BBa_K1951001+ RBS BBa_K1951002 by SpeI and PstI
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* Ligation of the 2 previous digestions in pSB1C3 to make the final BBa_K1951011 (E/P)
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* Transformation of the previous ligation in competant Tg1
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 +
</div>
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<div id="2016-09-09" class="evday">
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===== Friday, September 9th =====
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 +
* Derivatization of the sample cadA mutant complemented with BBa_K1951004 and cadA mutant complemented with BBa_K1951004+ RBS BBa_K1951001
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* Reisolement of the knockout fliC mutant W3110 from the transformation of the 09.08.2016
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* Miniprep of the BBa_K1951008 + BbsI 215-216 position and starters of BBa_K1951008 + BbsI 238-239 position
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* Digestion test by BBs1 and EcoRI using NEB buffer and electrophoresis
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* Colony PCR of BBa_K1951011 in pSB1C3 by digestion
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 +
 +
</div>
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<div id="2016-09-10" class="evday">
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===== Satursday, September 10th =====
 +
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* Swimming test of the knockout fliC mutant W3110 and the WT
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* Starters of the fliC mutant W3110
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 +
 +
</div>
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<div id="2016-09-11" class="evday">
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===== Sunday, September 11st =====
 +
 +
* Made competant of the fliC mutant W3110 strain 
 +
* Transformation of the previous competant with BBa_K1951008
 +
 +
 +
</div>
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<div id="2016-09-11" class="evday">
 +
 +
===== Sunday, September 11st =====
 +
 +
* Made competant of the fliC mutant W3110 strain 
 +
* Transformation of the previous competant with BBa_K1951008
 +
 +
 +
</div>
 
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Revision as of 21:19, 14 October 2016